TERMINATOR SEQUENCE-CONTAINING REVERSE PRIMER FOR OVEREXPRESSION AND LINEAR DNA

摘要

Plasmid vectors have been widely used as a carrier of a DNA sequence capable of expressing a target RNA in cells. However, construction of these plasmid vectors requires technical skill and time. Thus, a quicker and easier method is required therefor. To solve this problem, a method using a linear DNA that has been amplified by the PCR method is examined. However, this method is disadvantageous in that RNA expression in cells is extremely low. Under these circumstances, the present inventors attempted to develop an RNA expression method using a linear DNA which can be produced mainly by using the PCR method alone and which enables a high level of RNA expression. As the results of intensive studies on terminator sequences to be used in a linear DNA, the present inventors found a smallest unit of a terminator sequence enabling linear DNA expression equivalent to that when using a plasmid vector. A linear DNA including the aforesaid terminator sequence can be produced quickly and easily, and enables RNA expression at a higher level. The present invention has been completed based on these findings.

主权项

1. A reverse primer for RNA expression in cells using linear DNA, the reverse primer being composed of a complementary sequence of a DNA sequence that includes a terminator sequence, wherein the terminator sequence is composed of between 30 and 200 nucleobases, includes a sequence of nine connected nucleobases (A/T/G), (A/T/G), T, A, A, A, (A/T/G/C), (A/T/G/C), (A/G/C), and is derived from a terminator sequence of β-globin or SV40 (simian virus 40).